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The obtain optimum production conditions for Glucose Oxidase biosensor using software Qualtek-4 The Glucose Oxidase (Gox) electrochemical sensors were used to detect a little amount of glucose. In this paper, the pure platinum plate (length of 5 cm and a width of 3 mm), the Ag/AgCl electrode and platinum electrode (diameter of 1 mm) were applied as a working electrode (W.E), reference electrode and auxiliary electrode sequential. Factors measured, glucose concentration and pH is Phosphate-buffered saline (PBS) that design software QUALTEK-4 has been tested. The experiments performed tests of the software, with increasing concentration, the current output is increased. Optimal conditions were optioned in neutral pH and maximum glucose concentrations. After confirmation tests in optimum conditions the expected error rate of application lower than 10 %. It shows the true error rate test with high sensitivity and accuracy